Serum Protein Electrophoresis with Immunofixation. Dr.Ajay Phadke Centre Head SRL Diagnostics- Dr.Avinash Phadke’s Lab. What is electrophoresis?. Electrophoresis is a method of separating proteins based on their physical properties .

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This buffer flow is termed electro-osmosis or endosmosis, which also carries the proteins with it to some extent by mechanical flow, not by charge.

The actual distance traveled by a particular protein migrating in an electrical field is determined by the elec- tromotive force (a feature of the protein itself and the pH) and the electro- osmotic force (a function primarily of the support medium).

When the electro-osmotic force is greater than the electrophoretic force acting on weakly anionic proteins (e.g., γ-globulins), those proteins move from the application point toward the cathode, even though their charge is slightly negative.

Albumin concentrations are vital to the understanding and interpretation of calcium and magnesium levels because these ions are bound to albumin, and so decreases in albumin are directly responsible for depression of their concentrations

Principle: Apply the patients sample on gel. Separate the sample. Add antibody . If positive = on washing this sample remains because of large size of complex.

Immunotyping : similar principle. Automated, not labour intensive.

BASIC DIFFERENCE: way how sample is processed. WE MIX sample with antibody before processing.Complex is made EVEN BEFORE SEPERATION TAKES PLACE. Then injected into capillary. Monoclonal complex will MIGRATE SLOWLY and will NOT form a peak.

THEREFORE, in IF you are looking for the band to be PRESENT. While in IT you are looking for it to be ABSENT!

“In the context of screening, the serum FLC assay in combination with serum protein electrophoresis (PEL) and immunofixation yields high sensitivity, and negates the need for 24-h urine studies for diagnoses other than light chain amyloidosis (AL).”

• “...once diagnosis of a plasma cell disorder is made, 24-h urine studies are required for all patients.”

• “For AL screening, however, the urine IFE should still be done in addition to the serum tests including the serum FLC.”